scholarly journals Fabrication, characterization, and optimization of a novel copper-incorporated chitosan/gelatin-based scaffold for bone tissue engineering applications

Bioimpacts ◽  
2021 ◽  
Author(s):  
Azam Bozorgi ◽  
Masoud Mozafari ◽  
Mozafar Khazaei ◽  
Mansooreh Soleimani ◽  
Zahra Jamalpoor

Introduction: Fabricating composite scaffolds with improved physicochemical properties as artificial microenvironments are of great interest in bone tissue engineering. Given advantageous properties of nano-hydroxyapatite/chitosan/gelatin (nHA/Cs/Gel) scaffolds, the present study aimed to synthesize a modified nHA/Cs/Gel biomimetic scaffold with improved features. Methods: Pure and copper (Cu)-substituted nHA was synthesized using the chemical precipitation method under controlled pH and temperature. Pure and Cu-substituted nHA/Cs/Gel scaffolds were fabricated by salt-leaching/freeze-drying method. Physicochemical characteristics of nanoparticles and scaffolds were explored using XRD, FTIR, FE-SEM/EDX, and ICP. Besides, scaffold mechanical strength, degradation, porosity, swelling, biomineralization, and cytocompatibility were assessed. Results: Pure and Cu-substituted nHA were synthesized and characterized with appropriate Cu substitution and improved physical properties. All scaffolds were highly porous (porosity >98%) and Cu incorporation reduced porosity from 99.555 ± 0.394% to 98.69 ± 0.80% while enlarged the pore size to more than100 µm. Cu-substitution improved the scaffold mechanical strength and the best result was observed in nHA.Cu5%/Cs/Gel scaffolds by the compressive strength 88.869 ± 19.574 MPa. Furthermore, 3% and 5% Cu-substituted nHA enhanced the scaffold structural stability and supported osteoblast spread, adhesion, survival, mineralization, and proliferation. Moreover, long-term and sustainable Cu release from scaffolds was observed within 28 days. Conclusion: Cu-substituted nHA/Cs/Gel scaffolds mimic the porous structure and mechanical strength of cancellous bone, along with prolonged degradation and Cu release, osteoblast attachment, viability, calcium deposition, and proliferation. Taken together, our results indicate the upgraded properties of nHA.Cu5%/Cs/Gel scaffolds for future applications in bone tissue engineering.

2013 ◽  
Vol 849 ◽  
pp. 151-156 ◽  
Author(s):  
Tarin Sukhachiradet ◽  
Wassanai Wattanutchariya

Autograft is a general method used in orthopedic surgery for a bone replacement. However, the disadvantage of this method is the amount of risk factor to the donor sites. Currently, bone tissue engineering is another technique that could be implemented to solve this problem. Artificial bone scaffold generated by bone tissue engineering can be employed in order to accelerate damaged bone regeneration. In fact, this scaffold can be fabricated from synthetic contents such as bioceramics, biopolymers or composite. Three types of biomaterials: Chitosan, Hydroxyapatite (HA) and Fibroin were used to form porous scaffold. This research investigated the preparation of Hydroxyapatite and Fibroin from natural materials. Hydroxyapatite was synthesized from mollusk shell by wet chemical precipitation method. While, Fibroin was extracted from silk worms cocoons. Freeze drying method was employed to fabricate this composite porous scaffold. A mixing ratio of 1:2:1 among Fibroin: Chitosan: HA was studied to evaluate biodegradability, biocompatibility, porosity and pore structure of the output scaffolds. Results show that the output scaffolds have an interconnected porous structure with a pore size around 150-200μm and an average porosity of 94.26%. While the average degradation rate of the scaffold in lysozyme was 10.46% at 7 days. In addition, the biocompatibility test based on XTT assay test, shown that the scaffolds were non-cytotoxicity, which could be good for bone filling application in the future.


2016 ◽  
Vol 705 ◽  
pp. 315-319
Author(s):  
Wassanai Wattanutchariya ◽  
Anirut Chaijaruwanich ◽  
Tarin Sukhachiradet

Autografting is a bone replacement technique used in orthopedic surgery. Bone tissue engineering is a new technique that offers promise, and could help alleviate this risk. Bioceramics, biopolymers or composite can be fabricated for artificial bone scaffold and used for bone regeneration. This study used three types of biomaterials – hydroxyapatite (HA), fibroin, and chitosan – to form porous scaffold. HA and fibroin were prepared from natural materials. HA was synthesized from mollusk shell by wet chemical precipitation method, while silk fibroin was extracted from silk worm’s cocoons. The HA and fibroin were mixed in a variety of ratios along with a fixed amount of chitosan before fabricating composite porous scaffolds by freeze-drying. The resulting scaffolds were evaluated for biodegradability, biocompatibility, porosity pore morphology and mechanical property. The fabricated scaffolds had an interconnected porous structure with a pore size of 200-400 μm and porosity in a range of 93-95%. The average degradation rate of the scaffold in lysozyme was between 7-17% at 7 days. A biocompatibility test showed that the scaffold was non-cytotoxic, making it a good candidate for future bone tissue engineering applications.


2021 ◽  
Vol 25 (1) ◽  
Author(s):  
Mona Sari ◽  
Puspa Hening ◽  
Chotimah ◽  
Ika Dewi Ana ◽  
Yusril Yusuf

Abstract Background The application of bioceramic hydroxyapatite (HA) derived from materials high in calcium to tissue engineering has been of concern, namely scaffold. Scaffold pores allow for cell mobility metabolic processes, and delivery of oxygen and nutrients by blood vessel. Thus, pore architecture affects cell seeding efficiency, cell viability, migration, morphology, cell proliferation, cell differentiation, angiogenesis, mechanical strength of scaffolds, and, eventually, bone formation. Therefore, to improve the efficacy of bone regeneration, several important parameters of the pore architecture of scaffolds must be carefully controlled, including pore size, geometry, orientation, uniformity, interconnectivity, and porosity, which are interrelated and whose coordination affects the effectiveness of bone tissue engineering. The honeycomb (HCB) as natural polymeric porogen is used to pore forming agent of scaffolds. It is unique for fully interconnected and oriented pores of uniform size and high mechanical strength in the direction of the pores. The aim of this study was therefore to evaluate the effect of HCB concentration on macropore structure of the scaffolds. Methods Bioceramic hydroxyapatite (HA) was synthesized from abalone mussel shells (Halioitis asinina) using a precipitation method, and HA-based scaffolds were fabricated with honeycomb (HCB) as the porogen agent. Pore structure engineering was successfully carried out using HCB at concentrations of 10, 20, and 30 wt%. Results The Energy Dispersive X-Ray Spectroscopy (EDS) analysis revealed that the Ca/P molar ratio of HA was 1.67 (the stoichiometric ratio of HA). The Fourier Transform Infrared Spectroscopy (FTIR) spectra results for porous HA-based scaffolds and synthesized HA showed that no chemical decomposition occurred in the HA-based scaffold fabrication process. The porosity of the scaffold tended to increase when higher concentrations of HCB were added. XRD data show that the HCB was completely degraded from the scaffold material. The cell metabolic activity and morphology of the HA + HCB 30 wt% scaffold enable it to facilitate the attachment of MC3T3E1 cells on its surface. Conclusion HCB 30 wt% is the best concentration to fabricate the scaffold corresponding to the criteria for pores structure, crystallographic properties, chemical decomposition process and cell viability for bone tissue engineering.


2007 ◽  
Vol 5 (25) ◽  
pp. 899-907 ◽  
Author(s):  
M.A Wood ◽  
Y Yang ◽  
E Baas ◽  
D.O Meredith ◽  
R.G Richards ◽  
...  

A number of bone tissue engineering strategies use porous three-dimensional scaffolds in combination with bioreactor regimes. The ability to understand cell behaviour relative to strain profile will allow for the effects of mechanical conditioning in bone tissue engineering to be realized and optimized. We have designed a model system to investigate the effects of strain profile on bone cell behaviour. This simplified model has been designed with a view to providing insight into the types of strain distribution occurring across a single pore of a scaffold subjected to perfusion–compression conditioning. Local strains were calculated at the surface of the pore model using finite-element analysis. Scanning electron microscopy was used in secondary electron mode to identify cell morphology within the pore relative to local strains, while backscattered electron detection in combination with X-ray microanalysis was used to identify calcium deposition. Morphology was altered according to the level of strain experienced by bone cells, where cells subjected to compressive strains (up to 0.61%) appeared extremely rounded while those experiencing zero and tensile strain (up to 0.81%) were well spread. Osteoid mineralization was similarly shown to be dose dependent with respect to substrate strain within the pore model, with the highest level of calcium deposition identified in the intermediate zones of tension/compression.


2014 ◽  
Vol 2014 ◽  
pp. 1-6 ◽  
Author(s):  
Tae-Yeong Bak ◽  
Min-Suk Kook ◽  
Sang-Chul Jung ◽  
Byung-Hoon Kim

Porous polycaprolactone (PCL) scaffolds were fabricated by using the CO2gas foaming/salt leaching process and then PCL scaffolds surface was treated by oxygen or nitrogen gas plasma in order to enhance the cell adhesion, spreading, and proliferation. The PCL and NaCl were mixed in the ratios of 3 : 1. The supercritical CO2gas foaming process was carried out by solubilizing CO2within samples at 50°C and 8 MPa for 6 hr and depressurization rate was 0.4 MPa/s. The oxygen or nitrogen plasma treated porous PCL scaffolds were prepared at discharge power 100 W and 10 mTorr for 60 s. The mean pore size of porous PCL scaffolds showed 427.89 μm. The gas plasma treated porous PCL scaffolds surface showed hydrophilic property and the enhanced adhesion and proliferation of MC3T3-E1 cells comparing to untreated porous PCL scaffolds. The PCL scaffolds produced from the gas foaming/salt leaching and plasma surface treatment are suitable for potential applications in bone tissue engineering.


2020 ◽  
Vol 7 (1) ◽  
pp. 12 ◽  
Author(s):  
Robert Owen ◽  
Hossein Bahmaee ◽  
Frederik Claeyssens ◽  
Gwendolen C. Reilly

There is variability in the reported effects of compounds on osteoblasts arising from differences in experimental design and choice of cell type/origin. This makes it difficult to discern a compound’s action outside its original study and compare efficacy between compounds. Here, we investigated five compounds frequently reported as anabolic for osteoblasts (17β-estradiol (oestrogen), icariin, lactoferrin, lithium chloride, and menaquinone-4 (MK-4)) on human mesenchymal progenitors to assess their potential for bone tissue engineering with the aim of identifying a potential alternative to expensive recombinant growth factors such as bone morphogenetic protein 2 (BMP-2). Experiments were performed using the same culture conditions to allow direct comparison. The concentrations of compounds spanned two orders of magnitude to encompass the reported efficacious range and were applied continuously for 22 days. The effects on the proliferation (resazurin reduction and DNA quantification), osteogenic differentiation (alkaline phosphatase (ALP) activity), and mineralised matrix deposition (calcium and collagen quantification) were assessed. Of these compounds, only 10 µM MK-4 stimulated a significant anabolic response with 50% greater calcium deposition. Oestrogen and icariin had no significant effects, with the exception of 1 µM icariin, which increased the metabolic activity on days 8 and 22. 1000 µg/mL of lactoferrin and 10 mM lithium chloride both significantly reduced the mineralised matrix deposition in comparison to the vehicle control, despite the ALP activity being higher in lithium chloride-treated cells at day 15. This demonstrates that MK-4 is the most powerful stimulant of bone formation in hES-MPs of the compounds investigated, highlighting its potential in bone tissue engineering as a method of promoting bone formation, as well as its prospective use as an osteoporosis treatment.


2011 ◽  
Vol 493-494 ◽  
pp. 718-722 ◽  
Author(s):  
Masoud Hafezi-Ardakani ◽  
Faranak Kavian ◽  
Fatollah Moztarzadeh ◽  
Mohamadreza Baghaban Eslaminejad ◽  
Ali Zamanian ◽  
...  

A novel merwinite/ Poly(lactic-co-glycolic) nanocomposite was synthesized by a solvent casting/salt leaching technique with varying merwinite contents from 10 to 30% (w/w). Poly(lactic-co-glycolic) /merwinite foams with a co-continuous structure of interconnected pores were formed. The microstructure of the pores and the walls was controlled by varying the merwinite content. The pore structure becomes more and more irregular with increasing merwinite content. Pore sizes ranging from several microns to a few hundred microns were obtained. The degradation assessment of the scaffolds is performed in phosphate-buffered saline (PBS) solution at 37°C. Weight loss during storage at 37°C in PBS (pH 7.4) was determined for the scaffolds. Weight loss increased from pure to high content during incubation time. The prepared merwinite/ (Polylactic-co-glycolic) nanocomposite with uniform microstructure may be used in bone tissue engineering applications.


2005 ◽  
Vol 288-289 ◽  
pp. 79-82 ◽  
Author(s):  
Young Mee Jung ◽  
Soo Hyun Kim ◽  
Sang Soo Kim ◽  
Hee Jin You ◽  
Byoung Soo Kim ◽  
...  

A novel process was developed to fabricate polymer/ceramic composites for bone tissue engineering. The mixture of polylactic acid (PLA), calcium metaphosphate (CMP), and NaCl were compressed and subsequently heated. After dissolving the NaCl salts, porous biodegradable polymer/ceramic composite scaffolds were formed. The characteristics of the scaffolds were compared to those of scaffolds fabricated using a conventional solvent casting method, in terms of pore structure, pore size distribution, and mechanical properties. The scaffolds were seeded by osteoblasts and cultured in vitro or implanted into nude mice subcutaneously for up to 5 weeks. Cells were better grown to form tissue-like structures on CMP/PLA composites fabricated by the Press-and-Baking method. In addition, the alkaline phosphatase activity of and calcium deposition in the scaffolds explanted from mice were enhanced significantly for the scaffolds by Press-and-Baking compared to them by solvent casting. Taken together, these results suggest that CMP promote cell differentiation and proliferation via direct interaction with cells in the CMP/PLA composites. This novel PLA/CMP composite will be applicable for bone tissue engineering to support and cell differentiation and growth.


Author(s):  
Masud Rana Md. ◽  
Naznin Akhtar ◽  
Zahid Hasan Md. ◽  
Asaduzzaman S M

Bone tissue engineering with cells and synthetic extracellular matrix represents a new approach for the regeneration of mineralized tissue compared with the transplantation of bone. Hydroxyapatite (HA) and its composite with biopolymer are extensively developed and applied in bone tissue regeneration. The main aim of this study was to fabricate and characterize of HA apatite based biocompatible scaffold for bone tissue engineering. Scaffolds with different ratio of polymers (chitosan & alginate), and fixed amount of synthetic HA were prepared using in situ co precipitation method and mineral to polymer ratio was 1:1 to 1: 2 . A cross linker agent, 2-Hydroxylmethacrylate (HEMA) was added at different percentage (0.5-2%) into the selected composition and irradiated at 5- 25 kGy to optimize the proper mixing of components at the presence of HEMA. Fabricated scaffolds were analyzed to determine porosity, density, biodegradability, morphology and structural properties. Porosity and density of the prepared scaffold were 75 to 92% and 0.21 to 0.42 g/cm3 respectively. However, the swelling ratio of the fabricated scaffolds was ranged from 133 to 197%. Nonetheless, there had a reasonable in-vitro degradation of prepared scaffolds. Flourier transform infrared spectroscopy (FTIR) analysis showed intermolecular interaction between components in the scaffold. Pore size of scaffold was measured by scanning electron microscope and the value was 162-510 μm. It could be proposed that this scaffold fulfills all the main requirements to be considered as a bone substitute for biomedical application in near future.


2021 ◽  
Author(s):  
Chunhua Gong ◽  
Haixia Tang ◽  
Yanbo Wu ◽  
Juan Guo ◽  
Ruixue Guo ◽  
...  

Abstract In this paper, chitosan/hydroxyapatite (CS/n-HA) were synthesized by ultrasound-assisted precipitation combined with inverse crosslinking-emulsion method. In order to obtain a scaffold material with excellent properties, Calcium sulfate hemihydrate (CSH) were combined with CS-HA obtained CSH/CS/n-HA composite scaffold via setting citric acid as solidifying liquid, which possessed better biodegradability, bioactivity, mechanical properties. The physicochemical, morphological properties of scaffolds were characterized by FTIR, XRD and TFSEM. In addition, explored were the mechanical, degradable, biocompatibility and iron release properties. The mechanical strength study indicated that the compressive strength of the porous composite scaffold was influenced by adding an appropriate amount of CS/n-HA composite microspheres. It was proved that the composite scaffold with 6% CS/n-HA content obtained the highest mechanical strength (17.46±1.29 MPa). The results illustrated that the composite scaffold possessed biodegradability and can form hydroxyapatite by dynamic balance of Ca and P elements. The hemolysis tests demonstrate that materials are non-hemolytic and have good blood compatibility. Therefore, the developed composite scaffolds are safe medical inorganic materials, which can potentially be applied in bone tissue engineering.


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